The invention provides substituted phenyl compounds of general formula
wherein R1, T, U and Ar are as defined in the specification, a process for their preparation, pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions, and their use in therapy.
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1. A compound of formula
wherein:
each R1 independently represents a hydrogen or halogen atom, or a trifluoromethyl, cyano, nitro, c1-c6 alkyl or c1-c6 alkoxy group; T represents a group nh; U represents an oxygen or sulphur atom or a group nh; Ar represents a group
X represents a bond, an oxygen atom or a group CO, CH2, CH2O O(CH2)m, CH2OCH2, NR5, CH2NR5, NR5CH2, CH2NR5CH2, CONR5, S(O)n or SO2NR5; m is 1, 2 or 3; n is 0, 1 or 2; one of R2 and R3 represents a halogen, cyano, nitro, amino, hydroxyl, or a group selected from (i) c1-c6 alkyl optionally substituted by at least one c3-c6 cycloalkyl, (ii) c3-c8 cycloalkyl, (iii) c1-c6 alkyloxy optionally substituted by at least one c3-c6 cycloalkyl, (iv) c3-c8 cycloalkyloxy, (v) S(O)pc1-c6 alkyl, or (vi) S(O)qc3-c8 cycloalkyl, each of these groups being optionally substituted by one or more fluorine atoms, and the other of R2 and R3 represents a hydrogen or halogen atom or a methyl group; p is 0, 1 or 2; q is 0, 1 or 2; R4 represents a 3- to 9-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substitutents independently selected from fluorine atoms, hydroxyl, c1-c6 alkyl, acetyl, hydroxyC1-c6 alkyl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl; r is 1, 2, 3, 4, 5 or 6; R5 represents a hydrogen atom or a c1-c6 alkyl or c3-c8 cycloalkyl group; and R6 and R7 each independently represent a hydrogen atom or a c1-c6 alkyl or c3-c8 cycloalkyl group, or R6 and R7 together with the nitrogen atom to which they are attached to form a 3- to 8-membered saturated heterocyclic ring, provided that when R3 represents a cyano group, then X is other than a bond; or a pharmaceutically acceptable salt or solvate thereof.
3. A compound according to
5. A compound according to
(i) c1-c4 alkyl optionally substituted by at least one c5-c6 cycloalkyl, (ii) c5-c6 cycloalkyl, (iii) c1-c4 alkyloxy optionally substituted by at least one c5-c6 cycloalkyl, (iv) c5-c6 cycloalkyloxy, (v) S(O)pc1-c4 alkyl, or (vi) S(O)qc5-c6 cycloalkyl, each of these groups being optionally substituted by one to four fluorine atoms, and the other of R2 and R3 represents a hydrogen or halogen atom.
6. A compound according to
7. A compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, according to
2-(3,4-Difluorophenylamino)-N-(2-methyl-5-piperazin-1-ylmethyl-phenyl)-acetamide, trihydrochloride, 2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-piperazin-1-ylmethyl-phenyl)-thioacetamide, 2-(3,4-Difluoro-phenylamino)-N-(2-methyl-4-piperazin-1-ylmethyl-phenyl)-acetamide, trihydrochloride, N-(2-Chloro-5-piperazin-1-yl-phenyl)-2-(3,4-difluoro-phenylamino)-acetamide, trihydrochloride, (S)-2-(3,4-Difluoro-phenylamino)-N-[2-methyl-4-(2-pyrrolidin-1-ylmethyl-pyrrolidin-1-ylmethyl)-phenyl]-acetamide, trihydrochloride, 2-(3-Chloro-4-fluorophenylamino)-N-{2-methyl-5-[3-(4-methylpiperazin-1-yl)-propoxyl]phenyl}acetamide, (+/-)-2-(3-Chloro-4-fluoro-phenylamino)-N-[2-methyl-5-(piperidin-4-yloxy)-phenyl]-acetamide, dihydrochloride, 2-(3,4-Difluoro-phenylamino)-N-[2-methyl-4-(piperidin-4-yloxy)-phenyl]-acetamide, dihydrochloride, (±)N-[5-(3-Amino-pyrrolidin-1-yl)-2-methyl-phenyl]-2-(3,4-difluoro-phenylamino)-acetamide, trihydrochloride, 2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-piperazin-1-yl-phenyl)-acetamide, trihydrochloride, (S)-2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-(2-pyrrolidin-1-ylmethyl-pyrrolidin-1-ylmethyl)-phenyl)-acetamide, (S)-2-(3,4-Difluoro-phenylamino)-N-[5-(2-hydroxymethyl-pyrrolidin-1-ylmethyl)-2-methyl-phenyl]-acetamide, 2-(3,4-Difluoro-phenylamino)-N-[2-methyl-5-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-phenyl]-acetamide, 2-(3,4-Difluorophenylamino)-N-[2-methyl-3-(piperidin-4-yloxy)phenyl]acetamide trifluoroacetate, or N-[3-(4-Acetyl-piperazin-1-ylmethyl)-2-methylphenyl]-2-(3,4-difluorophenylamino)acetamide.
8. A process for the preparation of a compound of formula (I) as defined in
(i) when U represents an oxygen atom, X represents a CH2 group and R4 represents a 3- to 8-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substitutents independently selected from fluorine atoms, hydroxyl, c1-c6 alkyl, hydroxyC1-c6 alkyl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl, reacting a compound of formula
wherein one of R10 and R11 represents a hydrogen atom and the other of R10 and R11 represents a group --CH2L1 in which L1 represents a leaving group, and T, R1, R2, and R3 are as defined in formula (I), with a compound of formula in the presence of a base, wherein R4' represents a 3- to 8-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substitutents independently selected from fluorine atoms, hydroxyl, c1-c6 alkyl, hydroxyC1-c6 alkyl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl and wherein R6 and R7 are as defined in formula (I); or (ii) when U represents an oxygen atom and X represents an oxygen atom or a group O(CH2)m, reacting a compound of formula
wherein one of R12 and R13 represents a hydrogen atom and the other of R12 and R13 represents a hydroxyl group, and T, R1, R2, and R3 are as defined in formula (I), with a compound of formula wherein Y represents a bond or a group (CH2)m and m and R4 are as defined in formula (I), in the presence of 1,1-(azodicarbonyl)dipiperidine and tributylphosphine; or (iii) when U represents an oxygen atom and X represents a bond, an oxygen atom or a group O(CH2)m, NR5, NR5CH2, CO, CONR5, SO2 or SO2NR5 reacting a compound of formula
wherein one of R14 and R15 represents a group --X'--R4 and the other of R14 and R15 represents a hydrogen atom, X' represents a bond, an oxygen atom or a group O(CH2)m, NR5, NR5CH2, CO, CONR5, SO2 or SO2NR5, and m, R2, R3, R4 and R5 are as defined in formula (I), with a compound of formula
wherein T and R1 are as defined in formula (I), in the presence of a coupling reagent and a base; or (iv) when U represents an oxygen atom and X represents a bond or a group NR5 or NR5CH2, reacting a compound of formula
wherein one of R16 and R17 represents a leaving group, L2, and the other of R16 and R17 represents a hydrogen atom and T, R1, R2, and R3 are as defined in formula (I), with a compound of formula wherein Z represents a hydrogen atom or a group NHR5 or CH2NHR5 and R4 and R5 are as defined in formula (I), optionally in the presence of a palladium catalyst, a phosphine ligand and a base; or (v) when U represents an oxygen atom and X represents a group CH2O, reacting a compound of formula (II) as defined in (i) above with a compound of formula (V) as defined in (ii) above wherein Y represents a bond, in the presence of a base or in the presence of a metal salt; or (vi) when U represents an oxygen atom and X represents a group CH2NR5, reacting a compound of formula (II) as defined in (i) above with a compound of formula (IX) as defined in (iv) above wherein Z represents a group NHR5; or (vii) when U represents an oxygen atom and X represents a group CH2OCH2, reacting a compound of formula (II) as defined in (i) above with a compound of formula (V) as defined in (ii) above wherein Y represents a group CH2, in the presence of a base or in the presence of a metal salt; or (viii) when U represents an oxygen atom and X represents a group CH2NR5CH2, reacting a compound of formula (II) as defined in (i) above with a compound of formula (IX) as defined in (iv) above wherein Z represents a group CH2NHR5; or (ix) when U represents an oxygen atom, X represents a group CH2 and R4 represents an unsubstituted 4- to 6-membered saturated heterocyclic ring system containing one nitrogen atom, reacting a compound of formula (II) as defined in (i) above, with a compound of formula
wherein s and t independently represent 1 or 2; or (x) when U represents an oxygen atom and X represents a sulfur atom, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium and then with a compound of formula
wherein R4 is as defined in formula (I); or (xi) when U represents an oxygen atom and X represents a CH2 group, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium and then with a compound of formula
wherein R4 is as defined in formula (I), followed by a reduction reaction; or (xii) when U represents an oxygen atom and X represents a bond, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium and then with a compound of formula R4═O (XIII)
wherein R4 is as defined in formula (I), followed by a reduction reaction; or (xiii) when U represents a sulphur atom, reacting a corresponding compound of formula (I) in which U represents an oxygen atom with a thiolating agent; or (xiv) when U represents a group nh, reacting a corresponding compound of formula (I) in which U represents a sulphur atom with a suitable alkylating agent followed by reaction with ammonium chloride or ammonia; or (xv) when U represents an oxygen atom and X represents CONR5, reacting a compound of formula
wherein one of R' and R" represents a hydrogen atom and the other of R' and R" represents a carboxyl group and T, R1, R2 and R3 are as defined in formula (I), with a compound of formula (XIIIB), R4--NHR5, wherein R4 and R5 are as defined in formula (I); or (xvi) when U represents an oxygen atom, X represents CH2 and R4 is bonded to X through a nitrogen atom, reacting a compound of formula
wherein T, R1, R2 and R3 are as defined in formula (I), with methane sulphonyl chloride followed by reaction with a compound of formula (XIIID), R4"--H, wherein R4" is defined as for R4 in formula (I); and optionally after (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv), or (xvi) converting the compound of formula (I) to a further compound of formula (I) and/or forming a pharmaceutically acceptable salt or solvate of the compound of formula (I).
9. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as claimed in
10. A process for the preparation of a pharmaceutical composition as claimed in
11. A method of effecting immunosuppression which comprises administering to a patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as claimed in
12. A method of treating, or reducing the risk of, an obstructive airways disease in a patient suffering from, or at risk of, said disease, which comprises administering to a patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as claimed in
13. A method according to
14. A method of treating rheumatoid arthritis, which comprises administering to a patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as claimed in
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The present invention relates to substituted phenyl compounds, a process for their preparation, pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions, and their use in therapy.
The P2X7 receptor (previously known as P2Z receptor), which is a ligand-gated ion channel, is present on a variety of cell types, largely those known to be involved in the inflammatory/immune process, specifically, macrophages, mast cells and lymphocytes (T and B). Activation of the P2X7 receptor by extracellular nucleotides, in particular adenosine triphosphate, leads to the release of interleukin-1β (IL-1β) and giant cell formation (macrophages/microglial cells), degranulation (mast cells) and proliferation (T cells), apoptosis and L-selectin shedding (lymphocytes). P2X7 receptors are also located on antigen-presenting cells (APC), keratinocytes, salivary acinar cells (parotid cells), hepatocytes and mesangial cells.
Certain substituted acetamide compounds are known from EP-A-382 216 having anti-allergic activity.
It would be desirable to make compounds effective as P2X7 receptor antagonists for use in the treatment of inflammatory, immune or cardiovascular diseases, in the aetiologies of which the P2X7 receptor may play a role.
In accordance with the present invention, there is therefore provided a compound of general formula
wherein:
each R1 independently represents a hydrogen or halogen (e.g. fluorine, chlorine, bromine or iodine) atom, or a trifluoromethyl, cyano, nitro, C1-C6 alkyl or C1-C6 alkoxy group;
T represents an oxygen atom or, preferably, a group NH;
U represents an oxygen or sulphur atom or a group NH, preferably an oxygen or sulphur atom;
Ar represents a group
X represents a bond, an oxygen atom or a group CO, CH2, CH2O, O(CH2)m, CH2OCH2, NR5, CH2NR5, NR5CH2, CH2NR5CH2, CONR5, S(O)n or SO2NR5;
m is 1, 2or 3;
n is 0, 1 or 2;
one of R2 and R3 represents a halogen, cyano, nitro, amino, hydroxyl, or a group selected from C1-C6 alkyl optionally substituted by at least one C3-C6 cycloalkyl, C3-C8 cycloalkyl, C1-C6 alkyloxy optionally substituted by at least one C3-C6 cycloalkyl, C3-C8 cycloalkyloxy, S(O)pC1-C6 alkyl or S(O)qC3-C8 cycloalkyl, each of these groups being optionally substituted by one or more fluorine atoms, and the other of R2 and R3 represents a hydrogen or halogen atom or a methyl group;
p is 0, 1 or 2;
q is 0, 1 or 2;
R4 represents di(C1-2 alkyl)N(CH2)t where t is 0, 1 or 2 or imidazolyl, or R4 represents a 3- to 9-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substituents independently selected from fluorine atoms, hydroxyl, C1-C6 alkyl, acetyl, hydroxy C1-C6 alkyl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl, or R4 represents a 3- to 8-membered saturated carbocyclic ring system substituted by one or more substituents independently selected from NR6R7, --(CH2)rNR6R7 and --CONR6R7 the ring system being optionally further substituted by one or more substituents independently selected from fluorine atoms, hydroxyl and C1-C6 alkyl;
r is 1, 2, 3, 4, 5 or 6;
R5 represents a hydrogen atom or a C1-C6 alkyl or C3-C8 cycloalkyl group; and
R6 and R7 each independently represent a hydrogen atom or a C1-C6 alkyl or C3-C8 cycloalkyl group, or R6 and R7 together with the nitrogen atom to which they are attached form a 3- to 8-membered saturated heterocyclic ring, provided that when R3 represents a cyano group, then X is other than a bond;
or a pharmaceutically acceptable salt or solvate thereof.
In the context of the present specification, unless otherwise indicated, an alkyl substituent or alkyl moiety in a substituent group may be linear or branched. When one of R2 and R3 represents a C1-C6 alkyl/C1-C6 alkyloxy optionally substituted by at least one C3-C6 cycloalkyl, it should be understood that one or both of the alkyl and cycloalkyl moieties may be optionally substituted by fluorine atoms. A 3- to 9-membered saturated heterocyclic ring system containing one or two nitrogen atoms may be a monocyclic or bicyclic ring system. Similarly, a 3- to 8-membered saturated carbocyclic ring system may be a monocyclic or bicyclic ring system. The hydroxyl moiety in a hydroxyalkyl substituent group may be located in any suitable position in the alkyl group. Typically, the hydroxyl moiety will be located on a terminal carbon atom in a straight chain alkyl group. The alkyl groups in a dialkylamino moiety may be the same or different.
Preferably, at least one group R1 is other than a hydrogen atom, especially a halogen atom such as a fluorine or chlorine atom.
Preferably X represents a bond, an oxygen atom or a group CONH, CH2 or O(CH2)m.
One of R2 and R3 represents a halogen (e.g. fluorine, chlorine, bromine or iodine), cyano, nitro, amino, hydroxyl, or a group selected from C1-C6 alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl or hexyl) optionally substituted by at least one (e.g. 1, 2 or 3) C3-C6 cycloalkyl (i.e. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), C3-C8 cycloalkyl (e.g. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), C1-C6 alkyloxy (e.g. methoxy, ethoxy, isopropoxy or tert-butoxy) optionally substituted by at least one(e.g. 1, 2 or 3) C3-C6 cycloalkyl (i.e. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl), C3-C8 cycloalkyloxy (e.g. cyclopropyloxy, cyclobutyloxy, cyclopentyloxy or cyclohexyloxy), S(O)pC1-C6 alkyl (e.g. S(O)pmethyl, -ethyl, -propyl, -butyl, -pentyl or -hexyl) or S(O)qC3-C8 cycloalkyl (e.g. S(O)qcyclopropyl, -cyclobutyl, -cyclopentyl or -cyclohexyl), each of these groups being optionally substituted by one or more (e.g. 1, 2, 3 or 4) fluorine atoms, and the other of R2 and R3 represents a hydrogen or halogen (e.g. fluorine, chlorine, bromine or iodine) atom or a methyl group.
Preferably, one of R2 or R3 represents a halogen (especially chlorine) atom or a C1-C6 alkyl (especially methyl) group and the other of R2 or R3 represents a hydrogen atom.
In one aspect, R4 may represent a 3- to 9-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more (e.g. 1, 2, 3 or 4) substituents independently selected from fluorine atoms, hydroxyl, C1-C6 alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl or hexyl), acetyl, hydroxyC1-C6 alkyl (e.g. hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, hydroxypentyl or hydroxyhexyl), --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl.
The 3- to 9-membered saturated heterocyclic ring system in the group R4 may be a monocyclic ring system such as a pyrrolidinyl (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl or 3-pyrrolidinyl), piperidinyl (e.g. 1-piperidinyl, 2-piperidinyl, 3-piperidinyl or 4-piperidinyl), piperazinyl (e.g. 1-piperazinyl) or homopiperazinyl ring, or a bicyclic ring system such as
In another aspect, R4 may represent a 3- to 8-membered saturated carbocyclic ring system substituted by one or more (e.g. 1, 2 or 3) substituents independently selected from NR6R7, --(CH2)rNR6R7, and --CONR6R7, the ring system being optionally further substituted by one or more (e.g. 1, 2, 3 or 4) substituents independently selected from fluorine atoms, hydroxyl and C1-C6 alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl or hexyl).
The 3- to 8-membered saturated carbocyclic ring in the group R4 is preferably a monocyclic ring system such as a cyclopentyl or cyclohexyl ring.
Specific examples of groups R4 include:
When X represents a bond or a group CO, CH2 or SO2, R4 preferably represents a group:
When X represents an oxygen or sulphur atom or a group CH2O O(CH2)m, CH2OCH2, NR5, CH2NR5, NR5CH2, CH2NR5CH2, CONR5, SO or SO2NR5, R4 preferably represents a group:
R5 represents a hydrogen atom, or a C1-C6, preferably C1-C4, alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl or hexyl) or C3-C8, preferably C3-C6, cycloalkyl (e.g. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl) group.
R6 and R7 each independently represent a hydrogen atom, or a C1-C6, preferably C1-C4, alkyl (e.g. methyl, ethyl, propyl, butyl, pentyl or hexyl) or C3-C8, preferably C3-C6, cycloalkyl (e.g. cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl) group, or R6 and R7 together with the nitrogen atom to which they are attached form a 3- to 8-membered saturated heterocyclic ring such as a pyrrolidinyl or piperidinyl ring.
Preferred compounds of the invention include:
2-(3,4-Difluorophenylamino)-N-(2-methyl-5-piperazin-1-ylmethyl-phenyl)-acetamide, trihydrochloride,
2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-piperazin-1-ylmethyl-phenyl)-thioacetamide,
2-(3,4-Difluoro-phenylamino)-N-(2-methyl-4-piperazin-1-ylmethyl-phenyl)-acetamide, trihydrochloride,
N-(2-Chloro-5-piperazin-1-yl-phenyl)-2-(3,4-difluoro-phenyl amino)-acetamide, trihydrochloride,
(S)-2-(3,4-Difluoro-phenylamino)-N-[2-methyl-4-(2-pyolidin-1-ylmethyl-pyrrolidin-1-ylmethyl)-phenyl]-acetamide, trihydrochloride,
2-(3-Chloro-4-fluorophenylamino)-N-{2-methyl-5-[3-(4-methylpiperazin-1-yl)-propoxy]phenyl}acetamide,
(+/-)-2-(3-Chloro-4-fluoro-phenylamino)-N-[2-methyl-5-(piperidin4-yloxy)-phenyl]-acetamide, dihydrochloride,
2-(3,4-Difluoro-phenylamino)-N-[2-methyl-4-(piperidin-4-yloxy)-phenyl]-acetamide, dihydrochloride,
(±)N-[5-(3-Amino-pyrrolidin-1-yl)-2-methyl-phenyl]-2-(3,4-difluoro-phenylamino)-acetamide, trihydrochloride,
2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-piperazin-1-yl-phenyl)-acetamide, trihydrochloride,
(S)-2-(3,4-Difluoro-phenylamino)-N-(2-methyl-5-(2-pyrrolidin-1-ylmethyl-pyrrolidin-1-ylmethyl)-phenyl)-acetamide,
(S)-2-(3,4-fluoro-phenylamino)-N-[5-(2-hydroxymethyl-pyrrolidin-1-ylmethyl)-2-methyl-phenyl]-acetamide,
2-(3,4-Difluoro-phenylamino)-N-[2-methyl-5-(4-pyrimidin-2-yl-piperazin-1-ylmethyl)-phenyl]-acetamide,
2-(3,4-Difluorophenylamino)-N-[2-methyl-3-(piperidin-4-yloxy)phenyl]acetamide trifluoroacetate,
3-[2-(3,4-Difluorophenylamino)acetylamino]-N-(2-dimethylaminoethyl)-2-methylbenzamide,
N-[3-(4-Acetyl-piperazin-1-ylmethyl)-2-methylphenyl]-2-(3,4-difluorophenylamino)acetamide,
2-(3,4-Difluorophenylamino)-N-(3-imidazol-1-ylmethyl-2-methylphenyl)acetamide, and
2-(3,4-Difluorophenylamino)-N-(3-dimethylaminomethyl-2-methylphenyl)acetamide.
The present invention further provides a process for the preparation of a compound of formula (I) as defined above which comprises:
(i) when U represents an oxygen atom, X represents a CH2 group and R4 represents a 3- to 8-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substituents independently selected from fluorine atoms, hydroxyl, C1-C6 alkyl, hydroxyC1-C6 alkyl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl, reacting a compound of general formula
wherein one of R10 and R11 represents a hydrogen atom and the other of R10 and R11 represents a group --CH2L1 in which L1 represents a leaving group (e.g. a halogen atom), and T, R1, R2 and R3 are as defined in formula (I), with a compound of general formula
in the presence of a base (e.g. diisopropylethylamine), wherein R4' represents a 3- to 8-membered saturated heterocyclic ring system containing one or two nitrogen atoms, the heterocyclic ring system being optionally substituted by one or more substituents independently selected from fluorine atoms, hydroxyl, C1-C6 alkyl, hydroxyC1-C6 alykl, --NR6R7, --(CH2)rNR6R7, --CONR6R7 and pyrimidinyl and wherein R6 and R7 are as defined in formula (I); or
(ii) when U represents an oxygen atom and X represents an oxygen atom or a group O(CH2)m, reacting a compound of general formula
wherein one of R12 and R13 represents a hydrogen atom and the other of R12 and R13 represents a hydroxyl group, and T, R1, R2 and R3 are as defined in formula (I), with a compound of general formula
wherein Y represents a bond or a group (CH2)m and m and R4 are as defined in formula (I), in the presence of 1,1-(azodicarbonyl)dipiperidine and tributylphosphine (under conditions of the Mitsunobu reaction: Tetrahedron Lett. (1993), 34, 1639); or
(iii) when U represents an oxygen atom and X represents a bond, an oxygen atom or a group O(CH2)m, NR5, NR5CH2, CO, CONR5, SO2 or SO2NR5 reacting a compound of general formula
wherein one of R14 and R15 represents a group --X'--R4 and the other of R14or R15 represents a hydrogen atom, X' represents a bond, an oxygen atom or a group O(CH2)m, NR5, NR5CH2, CO, CONR5, SO2 or SO2NR5, and m, R2, R3, R4 and R5 are as defined in formula (I), with a compound of general formula
wherein T and R1 are as defined in formula (I), in the presence of a coupling reagent such as isobutylchloroformate or bromo-tris-pyrrolidino-phosphonium hexafluorophosphate and a base (e.g. diisopropylarmine); or
(iv) when U represents an oxygen atom and X represents a bond or a group NR5 or NR5CH2, reacting a compound of general formula
wherein one of R16 or R17 represents a leaving group, L2, such as a halogen atom and the other of R16 or R17 represents a hydrogen atom and T, R1, R2 and R3 are as defined in formula (I), with a compound of general formula
wherein Z represents a hydrogen atom or a group NHR5 or CH2NHR5 and R4 and R5 are as defined in formula (I), optionally in the presence of a palladium catalyst (e.g. palladium acetate), a phosphine ligand (e.g. BINAP) and a base (e.g. cesium carbonate); or
(v) when U represents an oxygen atom and X represents a group CH2O reacting a compound of formula (II) as defined in (i) above with a compound of formula (V) as defined in (ii) above wherein Y represents a bond, in the presence of a base (e.g. sodium hydride) or in the presence of a metal salt (e.g. silver trifluoromethanesulfonate); or
(vi) when U represents an oxygen atom and X represents a group CH2NR5, reacting a compound of formula (II) as defined in (i) above with a compound of formula (IX) as defined in (iv) above wherein Z represents a group NHR5; or
(vii) when U represents an oxygen atom and X represents a group CH2OCH2, reacting a compound of formula (II) as defined in (i) above with a compound of formula (V) as defined in (ii) above wherein Y represents a group CH2, in the presence of a base (e.g. sodium hydride) or in the presence of a metal salt (e.g. silver trifluoromethanesulfonate); or
(viii) when U represents an oxygen atom and X represents a group CH2NR5CH2, reacting a compound of formula (II) as defined in (i) above with a compound of formula (IX) as defined in (iv) above wherein Z represents a group CH2NHR5; or
(ix) when U represents an oxygen atom, X represents a group CH2 and R4 represents an unsubstituted 4- to 6-membered saturated heterocyclic ring system containing one nitrogen atom, reacting a compound of formula (II) as defined in (i) above, with a compound of general formula
wherein s and t independently represent 1 or 2; or
(x) when U represents an oxygen atom and X represents a sulfur atom, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium (e.g. at -70°C C.) and then with a compound of general formula
wherein R4 is as defined in formula (I); or
(xi) when U represents an oxygen atom and X represents a CH2 group, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium (e.g. at -70°C C.) and then with a compound of general formula
wherein R4 is as defined in formula (I), followed by a reduction reaction, e.g. with triethylsilane and trifluoroacetic acid or by treatment with methyloxalylchloride and triethylamine followed by tributyltin hydride in the presence of azobisisobutyronitrile; or
(xii) when U represents an oxygen atom and X represents a bond, reacting a compound of formula (VIII) as defined in (iv) above, with n-butyllithium (e.g. at -70°C C.) and then with a compound of general formula
wherein R4 is as defined in formula (I), followed by a reduction reaction, e.g. with triethylsilane and trifluoroacetic acid or by treatment with methyloxalylchloride and triethylamine followed by tributyltin hydride in the presence of azobisisobutyronitrile; or
(xiii) when U represents a sulphur atom, reacting a corresponding compound of formula (I) in which U represents an oxygen atom with a thiolating agent (such as Lawessons' reagent) at a temperature, for example, in the range from 0°C to 100°C C.;
(xiv) when U represents a group NH, reacting a corresponding compound of formula (I) in which U represents a sulphur atom with a suitable alkylating agent (e.g. methyl iodide) followed by reaction with ammonium chloride or ammonia;
(xv) when U represents an oxygen atom and X represents CONR5, reacting a compound of general formula
wherein one of R' or R" represents a hydrogen atom and the other of R' or R" represents a carboxyl group and T, R1, R2 and R3 are as defined in formula (I), with a compound of general formula (XIIIB), R4--NHR5, wherein R4 and R5 are as defined in formula (I); or
(xvi) when U represents an oxygen atom, X represents CH2 and R4 is bonded to X through a nitrogen atom, reacting a compound of general formula
wherein T, R1, R2 and R3 are as defined in formula (I), with methane sulphonyl chloride followed by reaction with a compound of general formula (XIIID), R4"--H, wherein R4" is defined as for R4 in formula (I) other than:
di(C1-2 alkyl)N(CH2)t where t is 1 or 2, and
3- to 8-membered saturated carbocyclic ring system substituted by one or more substituents independently selected from NR6R7, --(CH2)rNR6R7 and --CONR6R7, the ring system being optionally further substituted by one or more substituents independently selected from fluorine atoms, hydroxyl and C1-C6 alkyl;
and optionally after (i), (ii), (iii), (iv), (v), (vi), (vii), (viii), (ix), (x), (xi), (xii), (xiii), (xiv), (xv) or (xvi) converting the compound of formula (I) to a further compound of formula (I) and/or forming a pharmaceutically acceptable salt or solvate of the compound of formula (I).
The processes of the invention may conveniently be carried out in a solvent, e.g. an organic solvent such as dichloromethane, tetrahydrofuran, dioxane, xylene or dimethylformamide, at a temperature, e.g. in the range from -78 to 200°C C., preferably in the range from 0 to 150°C C.
Compounds of formula (II) in which L1 represents, for example, a chlorine atom may be prepared by reacting a compound of general formula
wherein one of R18 or R19 represents a hydrogen atom and the other of R18 and R19 represents a protected benzylalcohol group (the protecting group used may, for example, be tertbutyldimethylsilyl) and R2 and R3 are as defined in formula (II), with a compound of formula (VII) as defined above, in the presence of a coupling reagent such as isobutylchloroformate or bromo-tris-pyrrolidino-phosphonium hexafluorophosphate and a base (e.g. diisopropylamine), followed by deprotection and reaction with methanesulphonyl chloride in the presence of a base such as diisopropylamine.
Compounds of formula (IV) may be prepared by reacting a compound of general formula
wherein R2, R3, R12 and R13 are as defined in formula (IV), with a compound of formula (VII) as defined above, in the presence of a coupling reagent such as isobutylchloroformate or bromo-tris-pyrrolidino-phosphonium hexafluorophosphate and a base (e.g. diisopropylamine)
Compounds of formula (VI) may conveniently be prepared by reacting a compound of general formula
wherein L3 represents a leaving group (e.g. a hydroxyl group) and R2, R3, R14 and R15 are as defined in formula (VI), with diphenylphosphoryl azide in the presence of a base such as triethylamine.
Compounds of formula (XVI) in which X represents a bond, an oxygen atom or a group O(CH2)m, NR5 or NR5CH2 can be prepared by reacting a compound of general formula
wherein R20 represents a hydrogen atom or a C1-C6 alkyl group, one of R21 and R22 represents a leaving group, L4, such as a halogen atom (e.g. bromine or iodine) or a trifluoromethanesulfonate group and the other of R21 or R22 represents a hydrogen atom, and R2 and R3 are as defined in formula (XVI), with a compound of general formula
wherein X" represents a bond, an oxygen atom or a group O(CH2)m, NR5 or NR5CH2 and R4 is as defined in formula (I), in the presence of a palladium catalyst (1996 J. Am. Chem. Soc., 7215-6; 1997 J. Am. Chem. Soc., 3395), followed by a hydrolysis reaction (e.g. with sodium hydroxide).
Compounds of formula (XVI) in which X represents CO, CONR5, SO2 or SO2NR5 can be prepared by reacting a compound of general formula
wherein R23 represents a hydrogen atom or a C1-C6 alkyl group, one of R24 and R25 represents a group COL5 or SO2L5 and the other of R24 or R25 represents a hydrogen atom, L5 represents a leaving group (e.g. a halogen atom) and R2 and R3 are as defined in formula (XVI), with a compound of formula (IX) in which Z represents a hydrogen atom or a group NHR5, in the presence of a base such as diisopropylethylamine and catalytic N,N-dimethylaminopyridine, followed by a hydrolysis reaction (e.g. using sodium hydroxide).
Compounds of formula (VII) may be conveniently prepared by reacting a compound of general formula
wherein R26 represents a hydroxyl group or a protected nitrogen atom (protected by, for example, a tertbutyloxycarbonyl group) and R1 is as defined in formula (I), with an alkylating agent (e.g. methyl bromoacetate) followed by a saponification reaction.
Compounds of formula (VIII) may be prepared in an analogous manner to compounds of formula (IV) using, instead of the intermediate compound of formula (XV), an intermediate compound of general formula
R2, R3, R16 and R17 are as defined in formula (VIII).
Compounds of formula (X) can be prepared as described in Syn. Lett. (1998) 379-380.
Compounds of formulae (XIIIA) and (XIIIC) may be prepared by processes analogous to those already described.
Compounds of formulae (III), (V), (IX), (XI), (XII), (XIII), (XIIIB), (XIIID), (XIV), (XV), (XVII), (XVIII), (XIX), (XX) and, (XXI) are either commercially available, are well known in the literature or may be prepared easily using known techniques.
Compounds of formula (I) can be converted into further compounds of formula (I) using standard procedures. For example, compounds of formula (I) in which one of R2 and R3 represents a nitro group can be converted to compounds of formula (I) in which one of R2 and R3 represents an amino group by reduction using iron powder and ammonium chloride in ethanol/water under reflux conditions. The latter compounds can in turn be converted into compounds of formula (I) in which one of R2 and R3 represents a halogen atom, e.g. chlorine, by diazotization (e.g. with sodium nitrite) and reaction with copper chloride. Compounds of formula (I) in which R6 or R7 represents a hydrogen atom can be converted to compounds of formula (I) in which R6 or R7 represents a C1-C6 alkyl, C3-C8 cycloalkyl or a 3- to 8-membered saturated heterocyclic ring by standard chemical procedures.
It will be appreciated by those skilled in the art that in the processes of the present invention certain functional groups such as hydroxyl or amino groups in the starting reagents or intermediate compounds may need to be protected by protecting groups. Thus, the preparation of the compounds of formula (I) may involve at a certain stage the removal of one or more protecting groups.
The protection and deprotection of functional groups is described in `Protective Groups in Organic Chemistry`, edited by J. W. F. McOmie, Plenum Press (1973) and `Protective Groups in Organic Synthesis`, 2nd edition, T. W. Greene and P. G. M. Wuts, Wiley-Interscience (1991).
The compounds of formula (I) above may be converted to a pharmaceutically acceptable salt or solvate thereof, preferably an acid addition salt such as a hydrochloride, hydrobromide, phosphate, acetate, fumarate, maleate, tartrate, citrate, oxalate, methanesulphonate or p-toluenesulphonate, or an alkali metal salt such as a sodium or potassium salt.
Certain compounds of formula (I) are capable of existing in stereoisomeric forms. It will be understood that the invention encompasses all geometric and optical isomers of the compounds of formula (I) and mixtures thereof including racemates. Tautomers and mixtures thereof also form an aspect of the present invention.
The compounds of the present invention are advantageous in that they possess pharmacological activity. They are therefore indicated as pharmaceuticals for use in the treatment of rheumatoid arthritis, osteoarthritis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease (COPD), hyperresponsiveness of the airway, septic shock, glomerulonephritis, irritable bowel disease, Crohn's disease, ulcerative colitis, atherosclerosis, growth and metastases of malignant cells, myoblastic leukaemia, diabetes, Alzheimer's disease, meningitis, osteoporosis, burn injury, ischaemic heart disease, stroke and varicose veins.
Accordingly, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined for use in therapy.
In another aspect, the invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined in the manufacture of a medicament for use in therapy.
In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.
The invention further provides a method of effecting immunosuppression (e.g. in the treatment of rheumatoid arthritis, irritable bowel disease, atherosclerosis or psoriasis) which comprises administering a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined to a patient.
The invention also provides a method of treating an obstructive airways disease (e.g. asthma or COPD) which comprises administering to a patient a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined to a patient.
For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired and the disorder indicated. The daily dosage of the compound of formula (I)/salt/solvate (active ingredient) may be in the range from 0.001 mg/kg to 30 mg/kg.
The compounds of formula (I) and pharmaceutically acceptable salts and solvates thereof may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the formula (I) compound/salt/solvate (active ingredient) is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99% w (percent by weight), more preferably from 0.10 to 70% w, of active ingredient, and, from 1 to 99.95% w, more preferably from 30 to 99.90% w, of a pharmaceutically acceptable adjuvant, diluent or carrier, all percentages by weight being based on total composition.
Thus, the present invention also provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined in association with a pharmaceutically acceptable adjuvant, diluent or carrier.
The invention further provides a process for the preparation of a pharmaceutical composition of the invention which comprises mixing a compound of formula (I), or a pharmaceutically acceptable salt or solvate thereof, as hereinbefore defined with a pharmaceutically acceptable adjuvant, diluent or carrier.
The pharmaceutical composition of the invention may be administered topically (e.g. to the lung and/or airways or to the skin) in the form of solutions, suspensions, heptafluoroalkane aerosols and dry powder formulations; or systemically, e.g. by oral administration in the form of tablets, capsules, syrups, powders or granules, or by parenteral administration in the form of solutions or suspensions, or by subcutaneous administration or by rectal administration in the form of suppositories or transdermally.
The present invention will now be further explained by reference to the following illustrative examples.
To a solution of (3,4-difluorophenyl)-carbamic acid tert-butyl ester (Tetrahedron, (1992), 48(35), 7373) (15.1 g) in tetrahydrofuran (120 ml) at 0°C C. was added potassium tert-butoxide (94 ml of a 1.0 M solution in tetrahydrofuran) dropwise. After stirring for 2 hr methyl bromoacetate (9.1 ml) was added dropwise and ice-bath removed and reaction mixture stirred at room temperature overnight. Reaction mixture concentrated under reduced pressure, diluted with ethyl acetate and washed with water and brine, dried (Na2SO4) and concentrated to leave the sub-title compound as a brown oil (18.0 g).
1H NMR (DMSO-d6) δ7.43-7.12 (3H, m), 4.33 (2H, s), 3.67 (3H, s), 1.37 (9H, s).
To a solution of [tert-butoxycarbonyl-(3,4-difluorophenyl)-amino]-acetic acid methyl ester (14.7 g) in tetrahydrofuran (100 ml) was added water (100 ml) and lithium hydroxide monohydrate (6.2 g) and reaction mixture stirred at room temperature over-night before removal of tetrahydrofuran at reduced pressure. Residue was acidified to pH 4 by addition of KHSO4 (10% solution in water) and extracted with ethyl acetate. Organic extracts combined, dried (Na2SO4) and concentrated to leave the sub-title compound as a gum (12.3 g).
1H NMR (DMSO-d6) δ12.82 (1H, s), 7.37 (2H, m), 7.14 (1H, m), 4.22 (2H, s), 1.38 (9H, s).
To a solution of 4-chloromethyl-1-methyl-2-nitro-benzene (2 g) and triethylamine (3 ml) in DMF (10 ml) was added piperazine-1-carboxylic acid tert-butyl ester (2.01 g) and the stirred reaction heated at 78°C C. for 15 hr. After cooling to room temperature, the reaction was poured into ethyl acetate/water and the organic layer was separated. The aqueous layer was extracted with ethyl acetate. The combined organic extracts were washed with water, aqueous KHSO4, aqueous K2CO3 and brine, dried (Na2SO4) and concentrated to leave the sub-title compound as a brown oil (3.5 g).
1H NMR (DMSO-d6) δ7.89 (1H, d), 7.56 (1H, dd), 7.46 (1H, d), 3.55 (2H, s), 3.30 (7H, m), 2.33 (4H, m), 1.39 (9H, s).
To crude 4-(4-methyl-3-nitro-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (3.5 g) in EtOH/water (60 ml, 1:1) was added iron powder (4 g) and solid NH4Cl (4 g) and the mixture heated to reflux temperature for 1.5 hr. After cooling to room temperature the reaction was filtered through Celite and the filter cake washed with further EtOH. The majority of the solvent was removed in vacuo before addition of ethyl acetate and aqueous K2CO3. The organic layer was separated, washed with brine, dried (Na2SO4) and concentrated, to leave an oil that was triturated with isohexanes to afford a pale brown solid (3 g).
1H NMR (DMSO-d6) δ6.83 (1H, d), 6.55 (1H, d), 6.38 (1H, dd), 4.75 (2H, s), 3.28 (4H, m), 2.26 (4H, t), 2.01 (3H, s), 1.38 (9H, s).
To a solution of [tert-butoxycarbonyl-(3,4-difluorophenyl)-amino]-acetic acid (0.200 g) in tetrahydrofuran (2 ml) at 0°C C. was added triethylamine (110 ul) followed by dropwise addition of isobutylchloroformate (100 ul). After stirring for 1 hr, triethylamine (150 ul) was added followed by 4-(3-amino-4-methyl-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (0.234 g) in tetrahydrofuran (2 ml), and the reaction warmed to room temperature and then heated at 58°C C. overnight. After cooling to room temperature, the reaction mixture was filtered through Celite, the filtrate collected and the solvent removed in vacuo. Purification by NPHPLC (0-5% MeOH in CH2Cl2) afforded the sub-title compound as a pale yellow solid (0.300 g).
1H NMR (DMSO-d6) δ9.43 (1H, brs), 7.42 (2H, m), 7.35 (1H, s), 7.21 (1H, m), 7.15 (1H, d, J=7.6 Hz), 7.01 (1H, d, J=7.6 Hz), 4.38 (2H, s), 3.42 (2H, s), 3.29 (4H, m), 2.29 (4H, m), 2.15 (3H, s), 1.39 (9H, s), 1.38 (9H, s).
To a solution of 4-(3-{2-[tert-butoxycarbonyl-(3,4-difluoro-phenyl)-amino]-acetylamino}-4-methyl-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (0.100 g) in MeOH (3 ml) was added HCl (2ml of 4M solution in dioxane). After standing for several hours, a solid was filtered off, washed with acetone and dried in vacuo. The solid was re-suspended in acetone, sonicated for 5 min filtered and dried to afford the title compound (0.043 g, 51%).
Melting point: 232-248 (dec)°CC.
MS (ESI) 375 (M+H)+ for free base.
1H NMR (DMSO-d6) δ9.54 (3H, brs), 7.68 (1H, m), 7.37 (1H, d), 7.29 (1H, d), 7.16 (1H, dd), 6.61 (1H, ddd), 6.41 (1H, m), 4.33 (2H, brs), 3.92 (2H, s), 3.43-3.11 (10 H, m) 2.17 (3H, s).
To a solution of 4-(3-{2-[tert-butoxycarbonyl-(3,4-difluoro-phenyl)-amino]-acetylamino}-4-methyl-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (160 mg) in toluene (4 ml) was added Lawesson's reagent (100 mg) and the reaction heated at 100°C C. for 2.5 hr. After cooling to room temperature, the solvent was removed in vacuo to afford a yellow powder. Purification by NPHPLC (0-5% MeOH in CH2Cl2) afforded 4-(3-{2-[tert-butoxycarbonyl-(3,4-difluoro-phenyl)-amino]-thioacetylamino}-4-methyl-benzyl)-piperazine-1-carboxylic acid tert-butyl ester (120 mg). This was taken up in CH2Cl2 (1 ml) and trifluoroacetic acid (0.63 ml) added. After 2 hr stirring the reaction was poured into aqueous K2CO3 and the organic layer was separated, washed with aqueous K2CO3 and brine, dried (Na2SO4) and concentrated to leave yellow crystals. They were purified further by reverse phase HPLC to afford the title compound (14 mg).
Melting point: 72-82 (dec)°CC.
MS (ESI) 391 (M+H)+ for free base.
1H NMR (DMSO-d6) δ7.25 (2H, m), 7.18 (2H, m), 6.63 (2H, m), 6.45 (1H, d), 4.26 (2H, d), 3.57 (4H, m), 2.75 (4H, t), 2.35 (4H, brs), 2.06 (3H, s), (no thioamide proton observed).
To a solution of (3-methyl-4-nitro-phenyl)-methanol (1.22 g) and imidazole (0.99 g) in N,N-dimethylformamide (5 ml) was added tert-butyldimethylsilyl chloride (0.99 g) and the solution stirred for 5 hours before being poured into ethyl acetate/water and the organic layer separated, washed with water and brine, dried (Na2SO4) and concentrated. Purification of the residue by silica gel chromatography (eluting with 10% ethyl acetate in iso-hexane) gave 1.5 g of a pale brown oil. This was dissolved in ethanol (8 ml), cooled to 0°C C., and aqueous CuSO4 (0.5 ml of a 2M solution) added. Sodium borohydride (1.32 g) was added in small portions and the reaction was warmed to room temperature. Further portions of aqueous CuSO4 (0.5 ml) were added each hour until reduction was complete. Ethyl acetate was added, and the organic layer separated, washed with water and brine, dried (Na2SO4) and concentrated, to leave the sub-title compound as an oil (1.3 g).
1H NMR (DMSO-d6) δ6.81 (2H, m), 6.54 (1H, d), 4.73 (2H, brs), 4.48 (2H, s), 2.03 (3H, s), 0.97 (9H, s), 0.04 (6H, s).
To a solution of [tert-butoxycarbonyl-(3,4-difluorophenyl)-amino]-acetic acid (5.74 g) in tetrahydrofuran (40 ml) at 0°C C. was added triethylamine (3.2 ml) followed by isobutylchloroformate (2.9 ml) dropwise. After stirring for lhr, triethylamine (3.2 ml) was added followed by 4-(tert-butyl-dimethyl-silanyloxymethyl)-2-methyl-phenylamine (5.0 g) in tetrahydrofuran (3 ml), and the reaction warmed to room temperature and then heated at 58°C C. overnight. After cooling to 0°C C., tetrabutylammonium fluoride (7.6 ml of a 1M solution in tetrahydrofuran) was added dropwise and the reaction warmed to room temperature. After 3 hrs, the solution was poured into ethyl acetate/water and the organic layer separated, washed with KHSO4 (10% solution in water), water and brine, dried (Na2SO4) and concentrated. Purification of the residue by silica gel chromatography (eluting with 3% methanol in dichloromethane) gave an orange solid. Recrystallisation from 2-propanol/isohexane gave the sub-title compound as white crystals (2.8 g).
Melting point: 134-136°C C.
MS (ESI) 307 (M+H)+
1H NMR (DMSO-d6) δ9.41 (1H, s), 7.49-7.39 (3H, m), 7.23 (1H, m), 7.12 (2H, m), 5.11 (1H, t), 4.43 (2H, d), 4.37 (2H, s), 2.16 (3H, s), 1.39 (9H, s).
To a solution of (3,4-difluoro-phenyl)-[(4-hydroxymethyl-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester (0.100 g) in dry tetrahydrofuran (1 ml) was added N,N-diisopropylethylamine (0.17 ml) followed by methanesulfonyl chloride (0.04 ml) in a single portion. After 2 hr piperazine-1-carboxylic acid tert-butyl ester (0.137 g) was added, and the reaction heated at 70°C C. for 10 hrs. After cooling to room temperature, the reaction was poured into ethyl acetate/water. The organic layer was separated, washed with water, aqueous KHSO4 and aqueous NaHCO3. The organic layer was dried (Na2SO4) and concentrated. Purification by NPHPLC (eluting with 0-25% methanol in dichloromethane) gave a brown solid (0.120 g). This was taken up in methanol (3 ml) and HCl (1 ml of a 4M solution in dioxane) added. After 24 hrs, the resulting crystals were filtered off, washed with acetone and dried in vacuo to give the title compound as a white powder (0.060 g).
Melting Point: 278 (dec)°CC.
MS (ESI) 375 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.50 (3H, s), 7.58 (1H, d), 7.46 (1H, s), 7.42 (1H, d), 7.16 (1H, q), 6.61 (1H, ddd), 6.40 (1H, d), 4.44 (3H, s), 4.31 (2H, s), 3.57 (6H, s), 3.19 (2h, s). 2.16 (3H, s).
Piperazine-1-carboxylic acid tert-butyl ester (106 mg), 3-bromo-6-chlorobenzoic acid ethyl ester (125 mg), cesium carbonate (220 mg), palladium acetate (5 mg) and R-BINAP (22 mg) were combined in toluene (2 ml) and heated at 100°C C. in a sealed vessel for 48 hr. The cooled reaction was loaded onto a silica column and eluted with iso-hexane/ethyl acetate (4:1) to give the subtitle product (170 mg).
MS (APCI+ve) 269/271 (M+H-(tert-butyloxycarbonyl))+
1H NMR (CDCl3) δ7.28-7.31 (2H, m), 6.94 (1H, dd), 4.39 (2H, q), 3.58 (4H, t), 3.13 (4H, t), 1.48 (9H, s), 1.40 (3H, t).
To a solution of 4-(4-chloro-3-ethoxycarbonyl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (2.2 g) in tetrahydrofuran (10 ml) and water (5 ml) was added lithium hydroxide monohydrate (370 mg). Reaction mixture was stirred at room temperature for 48 hr before concentration at reduced pressure to give the subtitle product (2.1 g).
MS (APCI+ve) 340/342 (M+H)+ for free acid
1H NMR (D2O) δ7.40 (1H, d), 7.06-7.11 (2H, m), 3.65 (4H, t), 3.17 (4H, t), 1.50 (1H, s),
To a solution of 5-(4-tert-butoxycarbonyl-piperazin-1-yl)-2-chloro-benzoic acid lithium salt (2.1 g) in N,N-dimethylformamide (50 ml) was added diphenyl phosphorylazide (1.4 ml) and reaction mixture stirred at ambient temperature for 1 hour. Water (30 ml) was added and the reaction mixture heated to 60°C C. for 2 hours. After cooling to room temperature, the reaction was poured into ethyl acetate/water, the organic layer separated, washed with water, aqueous NaHCO3 and brine. The organic layer was dried (MgSO4) and concentrated. Purification of the residue by silica gel chromatography (eluting ethyl acetate/iso-hexane (1:3 to 1:1) gave the subtitle product (1.5 g).
MS (APCI+ve) 312/314 (M+H)+
1H NMR (DMSO-d6) δ6.98 (1H, d), 6.35 (1H, d), 6.19 (1H, dd), 5.12 (2H, s), 3.44 (4H, t), 2.98 (4H, t), 1.41 (9H, s).
To a solution of 4-(3-amino-4-chloro-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (500 mg) and N,N-diisopropylethylamine (0.8 ml) in tetrahydrofuran (20 ml) cooled to 0°C C. was added 2-chloroacetyl chloride (0.2 ml). After 2 hrs, the reaction was poured into ethyl acetate/water. The organic layer was separated, washed with water, aqueous NaHCO3 and brine, dried (MgSO4) and concentrated. Purification of the residue by silica gel chromatography (eluting ethyl acetate/iso-hexane (1:3 to 1:2) gave the sub-title product (0.5 g).
MS (APCI+ve) 388/390/392 (M+H)+
1H NMR (CDCl3) δ8.81 (1H, brs), 8.05 (1H, d), 7.24 (1H, d), 6.63 (1H, dd), 4.22 (2H, s), 3.56 (4H, t), 3.14 (4H, t), 1.48 (9H, s).
Potassium iodide (10 mg), 3,4-difluoroaniline (0.4 ml), N,N-diisopropyl ethylamine (0.9 ml) and 4-[4-chloro-3-(2-chloro-acetylamino)-phenyl]-piperazine-1-carboxylic acid tert-butyl ester (500 mg) were combined in N,N-dimethylformamide (10 ml) and heated at 90°C C. for 24 hr. After cooling to room temperature, the reaction was poured into ethyl acetate/water. The organic layer was separated, washed with water (×3) and brine, dried (MgSO4) and concentrated. Purification of the residue by silica gel chromatography (eluting ethyl acetate/iso-hexane (1:2 to 1:1) gave the subtitle product (360 mg).
MS (APCI+ve) 481/483 (M+H)+
1H NMR (CDCl3) δ9.01 (1H, s), 8.13 (1H, d), 7.18 (1H, d), 7.00 (1H, q), 6.59 (1H, dd), 6.54 (1H, ddd), 6.38 (1H, brd), 4.38 (1H, t), 3.91 (2H, d), 3.57 (4H, t), 3.15 (4H, t), 1.48 (9H, s).
4M Hydrochloric acid in dioxane (3 ml) was added to a solution of 4-{4-chloro-3-[2-(3,4-difluoro-phenylamino)-acetylamino]-phenyl}-piperazine-1-carboxylic acid tert-butyl ester (340 mg) in ether (20 ml). After 24 hours the precipitated solid was collected by filtration and washed successively with dichloromethane, acetonitrile, ethyl acetate, then recrystalised from ethanol to give the title product as the dihydrochloride (60 mg).
Melting point: 240°C C. (dec).
MS (APCI+ve) 381/383 (M+H)+
1H NMR (DMSO-d6) δ9.44 (1H, s), 9.21 (2H, brs), 7.59 (1H, d), 7.33 (1H, d), 7.15 (1H, q), 6.82 (1H, dd), 6.63 (1H, ddd), 6.60 (2H, brs), 6.40 (1H, t), 3.91 (2H, s), 3.34 (4H, t), 3.20 (4H, t).
To a solution of (3,4-difluoro-phenyl)-[(4-hydroxymethyl-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester (0.200 g) from Example 3b) in dry 1-methyl-2-pyrrolidinone (2 ml) was added N,N-diisopropylethylamine (0.52 ml) followed by methanesulfonyl chloride (0.16 ml) in a single portion. After 3 hours the reaction was poured into ethyl acetate/water. The organic layer was separated and washed with aqueous KHSO4, aqueous NaHCO3 and brine. The organic layer was dried (Na2SO4) and concentrated to a pale red oil which was dissolved in 1-methyl-2-pyrrolidinone (8 ml) and N,N-diisopropylethylamine (0.52 ml) and (S)-(+)-1-(2-pyrrolidinylmethyl)-pyrrolidine (0.41 ml) were added, and the reaction heated at 95°C C. overnight. After cooling to room temperature, the reaction was poured into ethyl acetate/water. The organic layer was separated and washed with aqueous KHSO4, aqueous K2CO3 and brine, dried (Na2SO4) and concentrated to a pale red oil. Purification by NPHPLC (eluting with 0-25% methanol in dichloromethane) gave a white solid which was dissolved in methanol (2 ml) and HCl (1 ml of a 4M solution in dioxane) added. After 2 hours, the solvent was removed and the residue recrystalised from isopropanol/acetonitrile to give the title compound as a white powder (0.071 g).
Melting Point: 178 (dec)°CC.
MS (ESI+ve) 443 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.49 (1H, s), 7.55 (2H, m), 7,47 (1H, d), 7.16 (1H, q), 6.10 (1H, ddd), 6.40 (1H, d), 4.68 (1H, d), 4.13 (1H, dd), 3.92 (4H, m), 3.58 (3H, m), 3.20 (1H, m), 3.13 (1H, m), 2.98 (2H, m), 2.17 (3H, s), 2.00 (8H, m), NH's not observed.
To a solution of 2-methyl-5-methoxyaniline (4.7 g) in tetrahydrofuran (50 ml), under nitrogen, was added diisopropylethylamine (12.6 ml) followed by dropwise addition of chloroacetyl chloride (2.7 ml) and the mixture stirred under nitrogen for 27 hours. The solution was poured into 2M aqueous hydrochloric acid (150 ml), extracted into diethyl ether (3×150 ml), washed with brine, dried (MgSO4), and concentrated to give the sub-title compound as a brown oil (5.15 g).
1H NMR (CDCl3) δ8.24 (1H, s), 7.64-7.63 (1H, d), 7.11-7.08 (1H, d), 6.70-6.66 (1H, dd), 4.23 (2H, s), 3.80 (3H, s), 2.24 (3H, s).
2-Chloro-N-(5-methoxy-2-methylphenyl)-acetamide (3 g), 3-chloro4-fluoroaniline (7.56 g), diisopropylethylamine (13.8 ml), potassium iodide (0.005 g) and anhydrous tetrahydrofuran (20 ml) were heated together at 105°C C. in a sealed tube for 19 hours. The mixture was then cooled and concentrated. Purification of the residue by silica gel chromatography (eluting with 1:1 iso-hexane/ethyl acetate) gave the sub-title compound (3.93 g).
MS (APCI+ve) 323 (M+H)+
1H NMR (CDCl3) δ8.42 (1H, s), 7.71-7.70( 1H, d), 7.04-6.89 (2H, m), 6.76-6.47 (3H, m), 4.38 (1H, t), 3.92-3.90 (2H, d), 3.80 (3H, s), 2.00 (3H, s).
To a solution of 2-(3-chloro-4-fluorophenylamino)-N-(5-methoxy-2-methylphenyl)-acetamide (3.93 g) in anhydrous dichloromethane (450 ml) was stirred under nitrogen at -78°C C. was added a solution of boron tribrouide (1M in dichloromethane, 95 ml) dropwise. The mixture was then allowed to warm to room temperature and stirred under nitrogen for 20 hours before being poured into ice/water (1 l), stirred vigorously for 1 hour and extracted into ethyl acetate (3×500 ml), dried (MgSO4) and concentrated. Purification of the residue by silica gel chromatography (eluting with 3% methanol in dichloromethane) gave the sub-title compound (2.65 g).
MS (APCI+ve) 309 (M+H)+
1H NMR (DMSO-d6) δ9.18 (1H, s), 9.14 (1H, s), 7.18-7.12 (1H, t), 7.01 (1H, s), 6.96-6.94 (1H, d), 6.73-6.70 (1H, m), 6.61-6.57 (1H, m), 6.49-6.46 (1H, d), 6.35-6.31 (1H, t), 3.88-3.86 (2H, d), 2.00 (3H, s).
To a solution of 2-(3-chloro-4-fluoro-phenylamino)-N-(5-hydroxy-2-methyl-phenyl)-acetamide (0.400 g) and 4-methyl-1-piperazinepropanol (0.270 g) in tetrahydrofuran (20 ml) under a nitrogen atmosphere was added tributylphosphine (0.64 ml) and 1,1'-(azodicarbonyl)dipiperidine (0.656 g) and the mixture stirred at ambient temperature for 20 hours. The reaction mixture was poured into diethylether and filtered through Celite. After concentration, the residue was purified by NPHPLC (eluting with 75% ethanol in dichloromethane) to afford the title compound (0.334 g).
MS (APCI+ve) 449 (M+H)+
1H NMR (DMSO-d6) δ9.24 (1H, s), 7.18-7.05 (3H, m), 6.74-6.56 (3H, m), 6.35-6.31 (1H, t), 3.94-3.87 (4H, q), 2.49-2.26 (10H, m), 2.18 (3H, s), 2.05 (3H, s), 1.87-1.78 (2H, quintet).
To a solution of 2-(3-chloro-4-fluoro-phenylamino)-N-(5-hydroxy-2-methyl-phenyl)acetamide (0.10 g) and 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (Synlett. 1998, 4, 379) (0.163 g) in tetrahydrofuran (3 ml) was added tributylphosphine (0.20 ml) and 1,1'-(azodicarbonyl)dipiperidine (0.204 g) and the mixture heated at 60°C C. for 3 hours. Further 4-hydroxy-piperidine-1-carboxylic acid tert-butyl ester (0.163 g), tributylphosphine (0.20 ml) and 1,1'-(azodicarbonyl)dipiperidine (0.204 g) were then added and the heating continued. After 3 hours, the reaction was cooled, poured into diethyl ether and filtered through Celite. After concentration, the residue was purified by NPHPLC (eluting with 0-5% ethanol in dichloromethane) to afford a white solid that was dissolved in methanol (3 ml) and HCl (2 ml of a 4M solution in dioxane) added. After 24 hours, the crystals were filtered off, washed with dichloromethane and dried in vacuo to give the title compound as a white powder (0.080 g).
Melting Point: 179 (dec)°CC.
MS (APCI+ve) 392/394 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.33 (1H, s), 8.90 (1H, brs), 8.85 (1H, brs), 7.18-7.09 (3H, m), 6.74 (2H, m), 6.60 (1H, m), 5.53 (2H, brs), 4.55 (1H, m), 3.90 (2H, s), 3.17 (2H, m), 3.05 (2H, m), 2.06 (3H, s), 2.06 (2H, m), 1.80 (2H, m).
To a solution of 4-amino-2-methyl-phenol (1 g) and imidazole (1.66 g) in tetrahydrofuran (10 ml) was added tert-butyldimethylsilyl chloride (1.1 g) and the solution stirred for 15 hours before being poured into ethyl acetate/water and the organic layer separated, washed with water, aqueous K2CO3 and brine, dried (Na2SO4) and concentrated to a red oil. Purification of the residue by silica gel chromatography (eluting with 50% is ethyl acetate in isohexane) gave a pale red oil (1.5 g) which was used without further purification.
1H NMR (DMSO-d6) δ6.39-6.28 (3H, m), 4.27 (2H, s), 1.89 (3H, s), 0.82 (9H, s), 0.00 (6H, s).
Prepared according to Example 3b) using [tert-butoxycarbonyl-(3,4-difluorophenyl)-amino]-acetic acid (1.82 g) (from Example 1b) and 4-(tert-butyl-dimethyl-silanyloxy)-3-methyl-phenylamine (1.5 g). Purification by silica gel chromatography (eluting with 30% ethyl acetate in isohexane) followed by trituration with diethyl ether gave the sub-title compound as a white powder (1.47 g).
1H NMR (DMSO-d6) δ9.25 (1H, s), 9.22 (1H, s), 7.44 (2H, m), 7.21 (1H, m), 7.03 (1H, d), 6.56 (2H, m), 4.32 (2H, s), 2.08 (3H, s), 1.39 (9H, s).
Prepared according to Example 7 using (3,4-difluoro-phenyl)-[(4-hydroxy-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester (100 mg) to leave the title compound as a white powder (0.033 g).
Melting Point: 210 (dec)°CC.
MS (APCI+ve) 376 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.33 (1H, s), 8.68 (2H, brs), 7.23 (1H, m), 7.15 (1H, q), 6.81 (2H, m), 6.59 (1H, ddd), 6.41 (1H, m), 6.10 (2H, brs), 4.58 (1H, m), 3.85 (2H, s), 3.20 (2H, m), 3.05 (2H, m), 2.08 (3H, s), 2.06 (2H, m), 1.82 (2H, m).
4-Fluoro-2-methyl-1-nitrobenzene (1 g), pyrrolidin-3-ylcarbamic acid tert-butyl ester (1.2 g), potassium carbonate (1.79 g) and dimethyl sulfoxide (10 ml) were heated together at 80°C C. under nitrogen for 15 hours. The mixture was cooled, diluted with ethyl acetate (200 ml), washed with 2M aqueous hydrochloric acid (200 ml), dried (MgSO4), and concentrated. Purification of the residue by silica gel chromatography (eluting with 20% ethyl acetate in isohexane) gave the sub-title compound (1.744 g) as a yellow solid.
1H NMR (DMSO-d6) δ8.03-8.00 (1H, d), 7.28-7.21 (1H, br d), 6.51-6.47 (2H, m), 4.20-4.12 (1H, br m), 3.61-3.16 (4H, m), 2.56 (3H, s), 2.20-2.08 (1H, m), 1.98-1.85 (1H, m), 1.39 (9H, s).
[1-(3-Methyl-4-nitrophenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (1.744 g), iron powder (1.52 g), ammonium chloride (1.45 g), ethanol (50 ml) and water (50 ml) were heated to reflux temperature under nitrogen for 2 hours. The mixture was cooled and the iron filtered off. Water (200 ml) was added to the residue and the product extracted into ethyl acetate (3×200 ml), dried (MgSO4), and concentrated to give the sub-title compound (1.56 g).
1H NMR (CDCl3) δ6.65 (1H, br s), 6.38 (2H, br m), 4.80 (1H, m), 4.33 (2H, br m), 3.60-2.80 (5H, m), 2.31-2.17 (4H, m), 1.92-1.82 (1H, m), 1.45 (9H, br s).
Prepared according to the procedures for Examples 4d), 4e) & 4f), without characterisation of intermediates, [1-(3-Amino-4-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (750 mg) gave the title compound (200 mg) as the trihydrochloride.
MS (APCI+ve) 361 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.22 (1H, s), 8.33 (2H, brs), 7.10-7.20 (2H, m), 6.60 (1H, ddd), 6.37-6.40 (3H, m), 5.70 (2H, brs), 3.91 (1H, brs), 3.82 (2H, s), 3.40-3.51 (2H, m), 3.20-3.30 (2H, m), 2.24-2.36 (2H, m), 2.07 (3H, s).
4-Fluoro-2-methyl-1-nitrobenzene (2 g), piperazine-1-carboxylic acid tert-butyl ester (4.8 g), potassium carbonate (3.57 g) and dimethyl sulfoxide (20 ml) were heated together at 80°C C. under nitrogen for 15 hours. The mixture was then cooled, diluted with ethyl acetate (200 ml), washed with 2M aqueous hydrochloric acid (200 ml), dried (MgSO4), and concentrated to give the sub-title compound (4.05 g).
MS (APCI+ve) 321 (M)+
1H NMR (DMSO-d6) δ8.02-7.98 (1H, d), 6.89-6.86 (2H, m), 3.45 (8H, s), 2.55 (3H, s), 1.42 (9H, s).
4-(3-Methyl-4-nitrophenyl)-piperazine-1-carboxylic acid tert-butyl ester (2 g), iron powder (1.74 g), ammonium chloride (1.67 g), ethanol (50 ml) and water (50 ml) were refluxed together under nitrogen for 2 hours. The mixture was cooled and the iron was filtered off. Water (200 ml) was added to the residue and the product extracted into ethyl acetate (3×200 ml), dried (MgSO4), and concentrated to give the sub-title compound (1.20 g).
1H NMR (DMSO-d6) δ6.62-6.52 (3H, m), 4.38 (2H, s), 3.41 (4H, br s), 2.83 (4H, br s), 2.02 (3H, s), 1.41 (9H, s).
Prepared according to the procedures for Examples 4d), 4e) & 4f), without characterisation of intermediates, 4-(3-amino-4-methyl-phenyl)-piperazine-1-carboxylic acid tert-butyl ester (490 mg) gave the title compound (200 mg) as the trihydrochloride.
Melting Point: >230°C C.
MS (APCI+ve) 361 (M+H)+ for free base
1H NMR (DMSO-d6) δ9.32 (1H, s), 9.22 (2H, brs), 7.22 (1H, d), 7.14 (1H, q), 6.85 (1H, d), 6.80 (1H, dd), 6.62 (1H, ddd), 6.41 (1H, brd), 6.03 (2H, brs), 3.91 (2H, brs), 3.35 (4H, brs), 3.20 (4H, brs), 2.08 (3H, s).
Prepared according to the procedure of Example 3a) using (3-amino-4-methyl-phenyl)-methanol (1.61 g), afforded the sub-title compound as a pale brown oil (2.13 g).
1H NMR (DMSO-d6) δ6.85 (1H, d), 6.55 (1H, s), 6.39 (1H, d), 4.77 (2H, s), 4.52 (2H, s), 2.01 (3H, s), 0.89 (9H, s), 0.06 (6H, s).
Prepared according to the method of Example 3b), from 5-(tert-butyl-dimethyl-silanyloxymethyl)-2-methyl-phenylamine (2.0 g) and [tert-butoxycarbonyl-(3,4-difluorophenyl)-amino]-acetic acid (2.30 g) to give the sub-title product (1.9 g) as a white solid.
1H NMR (DMSO-d6) δ9.42 (1H, s), 7.49-7.32 (3H, m), 7.24 (1H, m), 7.15 (1H, d), 7.03 (1H, d), 5.13 (1H, t), 4.43 (1H, d), 4.38 (1H, s), 2.15 (3H, s), 1.39 (9H, s).
To a solution of (3,4-difluoro-phenyl)-[(5-hydroxymethyl-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester (0.203 g) in dry 1-methyl-2-pyrrolidinone (2 ml) was added N,N-diisopropylethylamine (0.52 ml) followed by methanesulfonyl chloride (0.14 ml) in a single portion. After 3 hours, the reaction was poured into diethylether/water. The organic layer was separated and washed sequentially with water, aqueous KHSO4 and aqueous NaHCO3. The organics were then dried (Na2SO4) and concentrated. The resulting residue was dissolved in 1-methyl-2-pyrrolidinone (5 ml) and N,N-diisopropylethylamine (0.17 ml) added. An aliquot (50 mL) was removed and added to (S)-(+)-1-(2-pyrrolidinylmethyl)-pyrrolidine (125 mL of a 0.2 M solution in 0.4 M N,N-diisopropylethylamine in 1-methyl-2-pyrrolidinone) and the reaction heated at 95°C C. for 24 hours. After cooling to room temperature, the volatiles were removed in vacuo. Methanol (100 mL) was added, and after dissolution of the mixture, HCl (150 mL of a 4 M solution in dioxane) was added and the reaction stirred for a further 16 hours. The volatiles were removed and dimethyl sulfoxide (500 mL) was added to give the title compound as a 10 mM solution. An aliquot (30 mL) of this solution was diluted with dimethyl sulfoxide/water (220 mL of a 1:1 mixture) was analysed by HPLC on a 20 mm×3.9 mm Waters Symmetry C8 column, eluting with 30%-95% acetonitrile/ammonium acetate.
MS (APCI+ve) 443 (M+H)+
Prepared according to the procedure of Example 11c) using (3,4-difluoro-phenyl)-[(5-hydroxymethyl-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester and (S)-(+)-2-pyrrolidinemethanol to give the title compound as a 10 mM solution in dimethyl sulfoxide. An aliquot (30 mL) of this solution was diluted with dimethyl sulfoxide/water (220 mL of a 1:1 mixture) was analysed by HPLC on a 20 mm×3.9 mm Waters Symmetry C8 column, eluting with 30%-95% acetonitrile/ammonium acetate.
MS (APCI+ve) 390 (M+H)+
Prepared according to the procedure of Example 11c) using (3,4-difluoro-phenyl)-[(5-hydroxymethyl-2-methyl-phenylcarbamoyl)-methyl]-carbamic acid tert-butyl ester and 1-(2-pyrimidyl)piperazine dihydrochloride to give the title compound as a 10 mM solution in dimethyl sulfoxide. An aliquot (30 mL) of this solution was diluted with dimethyl sulfoxide/water (220 mL of a 1:1 mixture) was analysed by HPLC on a 20 mm×3.9 mm Waters Symmetry C8 column, eluting with 30%-95% acetonitrile/ammonium acetate.
MS (APCI+ve) 453 (M+H)+
The subtitle compound was prepared from 3-amino-o-cresol (1.83 g) by the method of Example 8 step (a).
MS (APCI) 238 (M+H)
1H NMR (CDCl3) δ6.86 (1H,t), 6.33 (1H,d), 6.28 (1H,d), 3.59 (2H, brs), 2.04 (3H,s), 1.01 (9H,s), 0.20 (6H,s).
The subtitle compound was prepared from the product of step (a) (3.25 g) by the method of example 4 step (d). Yield 4.2 g.
MS (APCI) 405 (M-H)
1H NMR (CDCl3) δ8.22 (1H, brs), 7.49 (1H,d), 7.09 (1H,t), 6.69 (1H,d), 4.23 (2H,s), 2.16 (3H,s), 1.02 (9H,s), 0.22 (6H,s).
The subtitle compound was prepared from the product of step (b) (4.2 g) by the method of Example 4 step (e). Yield 1.71 g.
MS (APCI) 405 (M-H)
1H NMR (CDCl3) δ8.38 (1H,s), 7.55 (1H,d), 7.10-6.99 (2H,m), 6.63 (1H,d), 6.54 (1H,m), 6.40 (1H, br d), 4.39 (1H,br t), 3.91 (2H,d), 1.94 (3H,s), 0.99 (9H,s), 0.19 (6H,s).
A solution of tetrabutylammonium fluoride in THF (1M, 4.6 ml) was added to a solution of the product from step (c) (1.71 g) in THF (20 ml). The mixture was stirred at room temperature for 1 hour and evaporated to dryness. The residue was purified by flash chromatography eluting with 3% ethanol (EtOH) in dichloromethane (CH2Cl2) to give the product as a white solid. Yield 0.82 g.
Melting point: 216°C C. (dec)
MS (APCI) 291 (M-H)
1H NMR (DMSO-d6) δ9.34 (1H,s), 9.27 (1H,s), 7.15 (1H,q), 6.94 (1H,m), 6.87 (1H,d), 6.64 (1H,d), 6.58 (1H,m), 6.39 (1H,br d), 6.30 (1H,t), 3.85 (2H,d), 1.92 (3H,s).
Tributylphosphine (0.55 ml) was added to a mixture of 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (0.44 g), and the product from step (d) (0.25 g) and 1,1'-(azodicarbonyl)dipiperidine (0.55 g) in tetrahydrofuran (10 ml). The mixture was heated at 60°C C. under nitrogen (N2) for 2.5 hours. A further 0.55 ml tributylphosphine, 0.44 g 4-hydroxypiperidine-1-carboxylic acid tert-butyl ester and 0.55 g 1,1'-(azodicarbonyl)dipiperidine were added and heating was continued for 3 hours. The mixture was cooled to room temperature, diluted with ether and filtered. The filtrate was concentrated in vacuo and the residue was purified by flash chromatography eluting with 1% EtOH in CH2Cl2. The product was dissolved in CH2Cl2 (10 ml) and treated with trifluoroacetic acid (10 ml). After 2 hours the reaction mixture was concentrated in vacuo and the residue was triturated with diethyl ether to give the trifluoroacetic acid salt of the product as a white solid. Yield 130 mg.
MS (APCI) 376 (M+H)
1H NMR (DMSO-d6) δ9.40 (1H,s), 8.55 (1H,br s), 8.44 (1H, br s), 7.20-7.09 (2H,m), 7.03 (1H,d), 6.88 (1H,d), 6.63-6.55 (1H,m), 6.40 (1H,d), 4.63 (1H,m), 3.87 (2H,s), 3.21 (2H,d), 3.11 (2H, br s), 2.05 (2H,m), 1.98 (3H,s), 1.83 (2H,m).
Trimethylsilyl chloride (5 ml) was added to a solution of 3-amino-2-methylbenzoic acid (3 g) in methanol (100 ml). The mixture was heated at reflux for 2 hours and concentrated in vacuo to give the hydrochloride salt. Yield 4.0 g.
1H NMR (DMSO-d6) δ7.60 (1H,d), 7.54 (1H,d), 7.35 (1H,t), 3.84 (3H,s), 2.42 (2H,s).
The subtitle compound was prepared from the product of step (a) (4.0 g) by the method of Example 4 step (d). Yield 4.8 g.
MS (APCI) 240 (M-1)
1H NMR (CDCl3) δ8.31 (1H,br s), 7.99 (1H,d), 7.71 (1H,d), 7.30 (1H,t), 4.26 (2H,s), 3.91 (3H,s), 2.51 (3H,s).
The title compound was prepared from the product of step (b) (6.68 g) by the method of Example 4 step (e). Yield 1.42 g.
1H NMR (DMSO-d6) δ9.59 (1H,s), 7.56 (1H,d), 7.53 (1H,d), 7.16 (1H,q), 6.60 (1H,m), 6.40 (1H,br d), 6.32 (1H,t), 3.90 (2H,d), 3.82 (3H,s), 2.26 (3H,s).
A mixture of the product from step (c) (1.33 g) and lithium hydroxide monohydrate (0.84 g) in methanol (120 ml) and water (25 ml) was heated at reflux for 2 hours, cooled and concentrated in vacuo. The residual aqueous solution was acidified with glacial acetic acid and extracted with ethyl acetate. The combined extracts were washed with water and brine, dried (MgSO4) and evaporated to give a cream solid. Yield 1.27 g.
Melting point: 198°C C. (dec)
MS (APCI) 319 (M-H)
1H NMR (DMSO-d6) δ12.92 (1H,br s), 9.55 (1H,s), 7.57 (1H,d), 7.49 (1H,d), 7.28 (1H,m), 7.16 (1H,q), 6.60 (1H,m), 6.40 (1H,br d), 6.32 (1H,t), 3.90 (2H,d), 2.28 (3H,s).
A mixture of 3-[2-(3,4-difluorophenylamino)acetylamino]-2-methylbenzoic acid (0.2 g), N,N-dimethylethylenediamine (0.08 ml), bromotripyrrolidinophosphonium hexafluorophosphate (PyBroP®) (0.34 g), dimethylaminopyridine (0.07 g) and N,N-diisopropylethylamine (0.31 ml) in dimethylformamide (5 ml) was stirred at room temperature under N2 for 18 hours. The reaction mixture was partitioned between ethyl acetate and water and the organic phase was washed with water and brine, dried (MgSO4) and evaporated. The crude product was triturated with diethyl ether. The resultant solid was triturated with ethyl acetate and filtered. Yield 70 mg.
MS (APCI) 391 (M+H)
1H NMR (DMSO-d6) δ9.44 (1H,s), 8.15 (1H,t), 7.42 (1H,d), 7.22-7.07 (3H,m), 6.63-6.56 (1H,m), 6.40 (1H,d), 6.31 (1H,t), 3.89 (2H,d), 3.28 (2H,m), 2.37 (2H,t), 2.17 (6H,s), 2.10 (3H,s).
N-[3-(4-Acetyl-piperazin-1-ylmethyl)-2-methylphenyl]-2-(3,4-difluorophenylamino)acetamide
The subtitle compound was prepared from 3-amino-2-methylbenzyl alcohol (2.0 g) by the method of Example 8 step (a).
1H NMR (CDCl3) δ7.01 (1H,t), 6.85 (1H,d), 6.64 (1H,d), 4.69 (2H,s), 3.59 (2H,brs), 2.08 (3H,s), 0.93 (9H,s), 0.10 (6H,s).
The subtitle compound was prepared from the product of step (a) (1.0 g) by the method of Example 4 step (d). Yield 1.3 g.
MS (APCI) 292 (M-Cl)
1H NMR (CDCl3) δ8.22 (1H,brs), 7.66 (1H,d), 7.32-7.21 (2H, m), 4.71 (2H,s), 2.20 (3H, s), 0.94 (9H,s), 0.01 (6H,s).
The subtitle compound was prepared from the product of step (b) (1.3 g) by the method of Example 4 step (e). Yield 1.03 g.
MS (APCI) 419 (M-H)
1H NMR (CDCl3) δ8.37 (1H,s), 7.71 (1H,d), 7.26-7.19 (2H,m), 7.01 (1H,q), 6.56-6.51 (1H,m), 6.40 (1H,m), 4.66 (2H,s), 3.92 (2H,d), 1.99 (3H,s), 0.91 (9H,s), 0.07 (6H,s).
The sub-title compound was prepared from the product of step (c) by the method of Example 14 step (d).
Melting point: 155°C C. (dec)
MS (APCI) 307 (M+H)
1H NMR (DMSO-d6) δ9.40 (1H,s), 7.24-7.12 (4H,m), 6.63-6.56 (1H,m), 6.42-6.39 (1H,m), 6.31 (1H,t), 5.08 (1H,t), 4.48 (2H,d), 3.87 (2H,d), 2.02 (3H,s).
Methane sulphonyl chloride (0.06ml) was added to a solution of the product of step (d) (0.12 g) and N,N-diisopropylethylamine (0.28 ml) in tetrahydrofuran (3ml). The reaction mixture was stirred at room temperature for 1.5 hours. A solution of 1-acetylpiperazine (0. 15 g) in tetrahydrofuran (1 ml) was added and the mixture was heated at reflux for 2 hours. The reaction mixture was partitioned between ethyl acetate and water and the organic phase was washed with water and brine, dried (MgSO4) and evaporated. The crude product was triturated with ethyl acetate/isohexane to give the product as a pale yellow solid.
Yield 0.1 g.
MS (APCI) 417 (M+H)
1H NMR (DMSO-d6) δ9.39 (1H,s), 7.28 (1H,d), 7.20-7.06 (3H,m), 6.60 (1H,m), 6.40 (1H,br d), 6.32 (1H,t), 3.88 (2H,d), 3.43 (2H,s), 3.37 (4H,m), 2.34 (2H,m), 2.29 (2H,m), 2.12 (3H,s), 1.97 (3H,s).
The title compound was prepared from imidazole by the method of Example 16.
Melting point: 160°C C. (dec)
MS (APCI) 357 (M+H)
1H NMR (DMSO-d6) δ9.47 (1H,s), 7.66 (1H,s), 7.30 (1H,d), 7.19-7.12 (2H,m), 7.07 (1H,s), 6.91 (1H,s), 6.78 (1H,d), 6.62-6.56 (1H,m), 6.35 (1H,d), 6.29 (1H,t), 5.23 (2H,s), 3.87 (2H,d), 2.05 (3H,s).
The title compound was prepared by the method of Example 16 using dimethylamine solution in tetrahydrofuran (2M).
Melting point:144°C C. (dec)
MS (APCI) 334 (M+H)
1H NMR (DMSO-d6) δ10.07 (1H,br s), 9.60 (1H,s), 7.45 (1H,d), 7.39 (1H,d), 7.27 (1H,m), 7.16 (1H,q), 6.64-6.59 (1H,m) 6.42 (1H,d), 4.33 (2H,d), 3.91 (2H,s), 2.73 (3H,s), 2.72 (3H,s), 2.18 (3H,s).
Pharmacological Analysis
Certain compounds such as benzoylbenzoyl adenosine triphosphate (bbATP) are known to be agonists of the P2X7 receptor, effecting the formation of pores in the plasma membrane (Drug Development Research (1996), 37(3), p.126). Consequently, when the receptor is activated using bbATP in the presence of ethidium bromide (a fluorescent DNA probe), an increase in the fluorescence of intracellular DNA-bound ethidium bromide is observed. The increase in fluorescence can be used as a measure of P2X7 receptor activation and therefore to quantify the effect of a compound on the P2X7 receptor.
In this manner, each of the title compounds of Examples 1 to 18 was tested for antagonist activity at the P2X7 receptor. Thus, the test was performed in 96-well flat bottomed microtiter plates, the wells being filled with 250 μl of test solution comprising 200 μl of a suspension of THP-1 cells (2.5×106 cells/ml) containing 10-4M ethidium bromide, 25 μl of a high potassium buffer solution containing 10-5M bbATP, and 25 μl of the high potassium buffer solution containing 3×10 M test compound. The plate was covered with a plastics sheet and incubated at 37°C C. for one hour. The plate was then read in a Perkin-Elmer fluorescent plate reader, excitation 520 nm, emission 595 nm, slit widths: Ex 15 nm, Em 20 nm. For the purposes of comparison, bbATP (a P2X7 receptor agonist) and pyridoxal 5-phosphate (a P2X7 receptor antagonist) were used separately in the test as controls. From the readings obtained, a pIC50 figure was calculated for each test compound, this figure being the negative logarithm of the concentration of test compound necessary to reduce the bbATP agonist activity by 50%. Each of the compounds of Examples 1 to 18 demonstrated antagonist activity, having a pIC50 figure >4.50.
Meghani, Premji, Furber, Mark, Thorne, Philip, Luker, Timothy J, Mortimore, Michael P
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